hrvatski

Glikozilacija izvanstaničnih proteina tijekom somatske embriogeneze bundeve (Cucurbita pepo L.)

Plant cells cultivated in a suspension secrete proteins in the cultivation medium, thereby affecting the further growth and cell differentiation. Glycoproteins make up the majority of extracellular proteins, many of which have structural and/or enzymatic role. Previous research has shown that the profile of extracted protein is changed in relation to changes during somatic embryogenesis induction. Moreover, an abnormal glycosylation may cause delay in plant embryo development. One of the best described glycoprotein associated with somatic embryogenesis of plants is endochitinase EP3, characteristic for the early stage of embryogenesis in carrot and Arabidopsis. The objective of this work was to compare the glycosylation status of proteins extracted in cell suspension of two pumpkin embriogenic lines induced by 2, 4-D or on a modified MS medium without hormones containing 1 mM NH4Cl as the sole source of nitrogen. It was noted that the cultivation medium with NH4Cl as the sole source of nitrogen undergo acidification. The addition of 25 mM MES stabilized pH of the culture medium, what enhanced the secretion of extracellular proteins. Presence of specific endochitinases and changes in their activity depended on the medium composition and developmental changes during somatic embryogenesis. Extracellular proteins were separated by SDS-PAGE electrophoresis, transferred to membrane and analyzed by processing membrane with different lectins (ConA, GNA, DSA, PNA). The results showed that there was a difference in the level of glycosylation caused by medium composition and pH.

C. pepo; dušik; endohitinaza; EP3; glikozilacija; izvanstanični proteini; pH; somatska embriogeneza

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